6-Bromoindirubin-3′-Oxime Regulates Colony Formation, Apoptosis, and Odonto/Osteogenic Differentiation in Human Dental Pulp Stem Cells

Abstract

6-bromoindirubin-3′-oxime (BIO) is a candidate small molecule that effectively modulates Wnt signalling owing to its stable property. The present study investigated the influence of BIO on the odonto/osteogenic differentiation of human dental pulp stem cells (hDPSCs). hDPSCs were treated with 200, 400, or 800 nM BIO, and the effects on hDPSC responses and osteogenic differentiation were assessed. BIO-mediated Wnt activation was confirmed by β-catenin nuclear translocation detected by immunofluorescence staining. BIO attenuated colony formation and cell migration determined by in vitro wound-healing assay. BIO increased early apoptotic cell population evaluated using flow cytometry. For osteogenic induction, BIO promoted alkaline phosphatase (ALP) activity and mineralisation in a dose-dependent manner. ALP, RUNX2, OCN, OSX, ANKH, DMP1, and DSPP mRNA expression were significantly upregulated. The OPG/RANKL expression ratio was also increased. Further, BIO attenuated adipogenic differentiation as demonstrated by decreased lipid accumulation and adipogenic-related gene expression. Bioinformatic analysis of RNA sequencing data from the BIO-treated hDPSCs revealed that BIO modulated pathways related to autophagy and actin cytoskeleton regulation. These findings demonstrated that BIO treatment promoted hDPSC osteogenic differentiation. Therefore, this small molecule is a strong candidate as a bioactive molecule to enhance dentin repair.

Human dental pulp stem cells derived extracellular matrix promotes mineralization via Hippo and Wnt pathways

“…Together with proteomic data, Wnt would be the promising pathway implicated in osteogenic differentiation regulated by dECM_DPSCs. Previous studies supported that Wnt activation using small molecules in Wnt agonists enhanced odonto/osteogenic differentiation of DPSCs 34,35 . The beneficial effect of Wnt on osteogenic differentiation was also reported in dental-related stem cells 36,37 .…”

Small molecule-mediated regenerative engineering for craniofacial and dentoalveolar bone

“…BIO was seen to increase β-catenin translocation which upregulated the Wnt signaling pathway in the DPSCs. BIO also increased expression of osteogenic genes ANKH, ENPP1, RUNX2, OSX, OCN, DSPP, DMP1, and OPG paired with downregulation of osteoclastogenic RANKL ( Kornsuthisopon et al, 2022 ).…”